中国兽医科学2011,Vol.41Issue(8):811-816,6.
卡介菌多糖核酸对泌乳奶牛外周血PMN吞噬功能及活性氧释放的影响
Effect of polysaccharide nucleic acid fraction of Bacillus Calmette Guerin on phagocytosis and generation of reactive oxygen species of peripheral blood neutrophil in lactating cow
摘要
Abstract
The aim of this study was to evaluate the effect of polysaccharide nucleic acid fraction of Bacillus Calmette Guerin (BCG-PSN) on phagocytosis and generation of reactive oxygen species(ROS) of lactating cow peripheral blood neutrophil(PMN) in vitro. Peripheral blood of healthy lactation cow at early stages of lactation was collected from the jugular vein by venipuncture in evacuated tubes containing heparin as anticoagulant. FITC-labeled E. coli 10μL was added to each sample after a 2 h co-incubation with different concentrations of BCG-PSN(final concentration of 0,2,5,10,40 and 80μg/mL). After further 20 min incubation at 37 ℃ ,phagocytosis was stopped by dipping into ice water. Then erythrocytes were lysed with lysing solution and the leucocytes were washed two times with PBS(0.01 mol/L,pH 7.2). The cells were then resuspended in PBS and analyzed on a FACScan flow cytometer. FL1 green channel captured FITC fluorescence. Phorbol myrisate acetate(PMA, final concentration of 100 ng/mL) was used to stimulate PMN after a 2 h co-incubation with different concentrations of BCG-PSN(final concentration of 0,2,5,10, 40 and 80μg/mL). Control tests of every group were set up with PBS. All tubes were incubated at 37 ℃ for 15 min followed with addition of working dihydrorhodamine 123 (DHR 123, final concentration of 5μmol/L) solution and incubated at 37 ℃ for 10 min. Then erythrocytes were lysed with lysing solution, washed two times with PBS(0.01 mol/L, pH 7.2) and stabilized by paraformaldehyde. The cells were re- suspended in PBS and analyzed on a FACScan flow cytometer. The results showed that BCG-PSN could significant reduced the generation of reactive oxygen species of PMN and had no effect on the phagocytosis of PMN. This suggests that BCG-PSN protects the mammary gland from inflammatory injury by attenuating the release of ROS but not by directly phagocytosis. BCG-PSN may be a potential candidate to prevent bovine mastitis.关键词
卡介菌多糖核酸/PMN/吞噬/活性氧/泌乳奶牛Key words
BCG-PSN, PMN/phagocytosis/ROS/lactating cow分类
农业科技引用本文复制引用
苗晋锋,顾蓓蓓,刘仪,邹思湘..卡介菌多糖核酸对泌乳奶牛外周血PMN吞噬功能及活性氧释放的影响[J].中国兽医科学,2011,41(8):811-816,6.基金项目
国家自然科学基金项目(31072104) ()
南京农业大学青年创新基金项目(Y200917) ()